Abstract
A new in vitro system is described for studying an interaction between Erwinia amylovoraand Pyrus communis (L.). The system uses single shoots placed onto the solid medium, and it enables to detect changes in pH of the medium and differential appearance of shoot necrosis. Shoots of susceptible cultivar (Williams) and tolerant cultivar (Harrow Sweet) were compared measuring the necrosis rate along the in vitroshoots and the pH variation following proton extrusion of both plant and pathogen. Shoots acidified differentially the culture medium depending on the presence of the pathogen, cultivar susceptibility and shoot inoculation methods. Differences in the tolerance level against pathogen among the cultivars were distinguishable only when the shoots were inoculated at the basal end. In susceptible cultivar, the necrosis appeared after 48 h of inoculation, while in tolerant cultivars after 72 h. This system is repeatable and more reliable than already known methods, such as in vitroleaf explants or in vivoplants; it can be used all around the year to test the gene expression and products essential to characterize the genes involved in the pathogenesis. This system showed the effects of E. amylovoraon the photosystem dependent system of host cells, confirmed by the effects of pathogen attack on the variation of chlorophyll a and chlorophyll b ratios and positive effects of light on the appearance of the first disease symptoms.
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References
Bauer DW & Beer SV (1991) Further characterization of an hrp gene cluster of Erwinia amylovora. Mol. Plant-Microbe Interact. 5: 493–499
Beer SV, Rundle JR & Wodzinski RS (1983) Interaction between Erwinia amylovora and Erwinia herbicola in vitro, in immature pear fruits and in apple blossoms. Acta Horticult. 151: 203–204
Bogdanove AJ, Wei ZM, Zhao L & Beer SV (1996) Erwinia amylovora secretes harpin via a type III pathway and contains a homolog of yopN of Yersinia spp. J. Bacteriol. 178: 1720–1730
Bogs J, Bruchmüller I, Erbar C & Geider K (1998) Colanization of host plants by the fireblight pathogen Erwinia amylovora marked with genes for bioluminescence and fluorescence. Phytopathology 88: 416–421
Brisset NM & Paulin JP (1992) A reliable strategy for the study of disease and hypersensitive reaction induced by Erwinia amylovora. Plant Sci. 85: 171–177
Gaudriault S, Malandrin L, Paulin JP & Barny MA (1997) DspA, an essential pathogenicity factor of Erwinia amylovora showing homology with AvrE of Pseudomonas syringae, is secreted via the Hrp secretion pathway in a DspB-dependent way. Mol. Microbiol. 26: 1057–1069
Genoud T, Buchala A, Chua NH & Métraux JP (2002) Phytochrome signalling modulates the SA-perceptive pathways in Arabidopsis. Plant J. 31: 80–95
Heldt HW (1997) Plant biochemistry and molecular biology. Oxford University Press, New York
Huang PY & Goodman RN (1976) Ultrastructural modifications in apple stems induced by Erwinia amylovora and the fire blight toxin. Phytopathology 66: 269–270
Jin Q, Hu W, Brown I, McGhee G, Hart P, Jones AL & He SY (2001) Visualisation of secreted Hrp and Avr proteins along the Hrp pilus during type III secretion in Erwinia amylovora and Pseudomonas syringae. Mol. Microbiol. 40: 1129–1139
Kurdjian A & Guem J (1989) Intracellular pH: measurement and importance in cell activity. Ann. Rev. Plant Physiol. Plant Mol. Biol. 40: 271–303
Leblay C, Chevreau E & Robin LM (1991) Adventitious shoot regeneration from in vitro leaves of several pear cultivar (Pyrus communis L.). Plant Cell Tiss. Org. Cult. 25: 99–105
Moran R (1982) Formulas for determination of chlorophyllous pigments extracted with N,N-dimethylformamide. Plant Physiol. 69: 1376–1381
Palmgren MG (2001) Plant plasma membrane H+-ATPase: powerhouses for nutrient uptake. Ann. Rev. Plant Physiol. Plant Mol. Biol. 52: 817–845
Pike SM, Adam AL, Pu XA, Hoyos ME, Laby R, Beer SV & Novacky A (1998) Effects of Erwinia amylovora harpin on tobacco leaf cell membranes are related to leaf necrosis and electrolyte leakage are distinct from perturbation caused by inoculation of E. amylovora. Physiol. Mol. Plant Pathol. 53: 39–60
Popham PL, Pike SM & Novacky A (1995) The effects of harpin from Erwinia amylovora on the plasma membrane of suspension-cultured tobacco cells. Physiol. Plant Pathol. 47: 39–50
Quoirin M, Lepoivre P & Boxus P (1977) Un premier bilan de dix années de recherche sur les cultures de méristèmes et la multiplication in vitro de fruitiers ligneux. Compte rendu des recherches, Station des Cultures Fruitières et Maraichères de Gembloux (1976–1977): 93–117
Rao MV, Paliyath G, Ormrod DP, Murr DP & Watkins CB (1997) Influence of salycylic acid on H2O2 production, oxidative stress, and H2O2-metabolizing enzymes. Plant Physiol. 115: 137–149
Samuilov VD, Lagunova EM, Dzyubinskaya EV, Izyumov DS, Kiselevsky DB & Makarova YV (2002) Involvement of chloroplasts in the programmed death of plant cells. Biochemistry (Moscow) 67: 627–634
Suhayda CG & Goodman RN (1981) Early proliferation and migration and subsequently xylem occlusion by Erwinia amylovora polysaccharide (EPS) in apple shoots. Phytopathology 71: 697–707
Vanneste JL (1995) Erwinia amylovora. In: Singh US, Singh RP & Kohmoto K (eds) Pathogenesis and Host Specificity in Plant Diseases: Histopathological, Biochemical, Genetic and Molecular Bases, Vol. 1. (pp. 21–46). Prokaryotes. Pergamon Press, Oxford, London
Verhoeven AS, Adams III WW, Denning-Adams B, Croce R & Bassi R (1999) Xanthophyll cycle pigment localization and dynamics during exposure to low temperatures and light stress in Vinca major. Plant Physiol. 120: 727–737
Wei ZM & Beer SV (1995) hrpL activates Erwinia amylovora hrp gene cluster transcription and is a member of the ECF subfamily of r factors. J. Bacteriol. 177: 6201–6210
Wei ZM, Kim JF & Beer SV (2000) Regulation of hrp genes and type III protein secretion in Erwinia amylovora by HrpX/HrpY, a novel two component system and Hrps. Mol. Plant-Microbe Interact. 13: 1251–1262
Wei ZM, Laby RJ, Zumoff CH, Bauer DW, He SY, Collmer A & Beer SV (1992) Harpin elicitor of the hypersensitive responses produced by the plant pathogen Erwinia amylovora. Science 257: 85–88
Xie Z & Chen Z (2000) Harpin-induced hypersensitive cell death is associated with altered mitochondrial function in tobacco cells. Mol. Plant-Microbe Interact. 13: 183–190
Zimmermann R, Iturriaga R & Becker-Birck J (1978) Simultaneous determination of the total number of acquatic bacteria and the number thereof involved in respiration. Appl. Environ. Microbiol. 36: 926–935
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Abdollahi, H., Rugini, E., Ruzzi, M. et al. In vitro system for studying the interaction between Erwinia amylovora and genotypes of pear. Plant Cell, Tissue and Organ Culture 79, 203–212 (2004). https://doi.org/10.1007/s11240-004-0661-0
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DOI: https://doi.org/10.1007/s11240-004-0661-0